Galpin/DA/N2 IIIBenv 
3.1 MOUSE MODEL 
3.1.1 CTL Induction By HIV-IT (V) 
The following murine studies employed the BALB/c syngeneic cell line 
BC10ME (BC) and the HIV-IT (V)-transduced BC10ME cell line (BCenv) as control 
and HIV-1 env-expressing CTL targets, respectively. BCenv transduced cells express 
the HIV-1 envelope and rev (env/rev) proteins, and the neo r selectable marker protein. 
Mice were immunized with one or two injections of HIV-IT (V), and the primed 
splenocytes were stimulated in vitro with irradiated BCenv cells. The in vitro 
stimulation is routinely used to increase the number of cytotoxic cells for optimal 
detection in the CTL assay. The resulting effector cells were tested for CTL activity 
by measuring lysis of radiolabeled BCenv and control BC target cells in a 4 - 5 hour 
chromium (-^Crj-release assay 48 * 49 * 50 . 
Examination of the accumulated data in the mouse model reveals variability in 
the ability to generate CTL responses to HIV-IT (V) (Figure 8). These data reveal no 
statistically significant differences in CTL induction with one or two injections, or 
over the range of vector doses administered (10 4 - 10 7 cfu). However, for a number of 
reasons, it is not surprising that considerable variability in CTL induction was 
observed in these experiments. Our experimental approach throughout these studies 
was to examine parameters which might impact CTL induction. Firstly, these data 
include CTL induction by fourteen different HIV-IT (V) lots and five different 
HIV-IT (V) formulations. Secondly, Figure 8 includes experiments designed to 
optimize CTL responses, examining such parameters as the route of injection, the 
number of administrations, the injection schedule and the volume of HIV-IT (V) 
injected. Finally, the induction of a specific CTL response is a result of interactions in 
a complex biological system where some variation in response is expected. 
48 Wamer, J.F., et al., AIDS Research & Human Retroviruses , 7:645-655 (1991). 
49 Wamer, J.F., et al.,7. Immunol., 122:575-581 (1979). 
-^Brunner, K.T., et al.. Immunology, 14:181 (1968). 
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